LOCUS       sequence-262935-        4401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4401)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4401)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221_SLC2A12
FEATURES             Location/Qualifiers
     source          1..4401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2526..2625)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2642..2661)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2643..2661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2666..2682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2666..2682)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2784..2803)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2795..3604
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2862..2881)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3751..4339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4240..4259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggtgc ctgtggagaa taccgagggc ccatccctgc tgaaccagaa
      721 gggcaccgcc gtggagacag agggaagcgg atccaggcac ccaccttggg ccagaggctg
      781 cggcatgttc acctttctga gctccgtgac agcagccgtg tccggactgc tggtgggata
      841 cgagctgggc atcatctctg gcgccctgct gcagatcaag accctgctgg ccctgagctg
      901 tcacgagcag gagatggtgg tgtctagcct ggtcatcggc gccctgctgg catccctgac
      961 cggaggcgtg ctgatcgaca gatatggccg gagaacagcc atcatcctgt cctcttgcct
     1021 gctgggcctg ggctctctgg tgctgatcct gtctctgagc tacaccgtgc tgatcgtggg
     1081 ccgcatcgcc atcggcgtgt ccatctctct gagctccatc gccacatgcg tgtacatcgc
     1141 agagatcgca cctcagcaca ggaggggact gctggtgagc ctgaatgagc tgatgatcgt
     1201 gatcggcatc ctgtccgcct acatctctaa ttatgccttc gccaacgtgt ttcacggctg
     1261 gaagtacatg ttcggactgg tcatcccact gggcgtgctg caggcaatcg ccatgtattt
     1321 cctgccaccc agccctcggt tcctggtcat gaagggacag gagggagcag catctaaggt
     1381 gctgggccgg ctgagagccc tgagcgacac cacagaggag ctgaccgtga tcaagtctag
     1441 cctgaaggat gagtaccagt atagcttctg ggacctgttt agatccaagg ataacatgag
     1501 gacccgcatc atgatcggcc tgacactggt gttctttgtg cagatcaccg gccagccaaa
     1561 tatcctgttc tacgcctcca cagtgctgaa gtctgtgggc tttcagagca acgaggcagc
     1621 atctctggca agcaccggag tgggcgtggt gaaagtgatc tctaccatcc cagccacact
     1681 gctggtggac cacgtgggca gcaagacatt tctgtgcatc ggctcctctg tgatggccgc
     1741 ctccctggtg accatgggca tcgtgaatct gaacatccac atgaatttca cacacatctg
     1801 taggtcccac aattctatca accagagcct ggacgagtcc gtgatctacg gccccggcaa
     1861 cctgtccacc aacaataaca cactgcggga tcactttaag ggcatcagct cccactctag
     1921 atctagcctg atgcctctga ggaatgacgt ggataagagg ggagagacca caagcgcctc
     1981 cctgctgaac gcaggactga gccacaccga gtaccagatc gtgacagacc caggcgatgt
     2041 gcccgccttc ctgaagtggc tgagcctggc ctccctgctg gtgtatgtgg ccgcctttag
     2101 catcggcctg ggccctatgc catggctggt gctgtccgag atcttcccag gaggaatcag
     2161 gggaagagca atggcactga cctcctctat gaattggggc atcaacctgc tgatctccct
     2221 gacattcctg accgtgacag atctgatcgg cctgccttgg gtgtgcttta tctacaccat
     2281 catgtctctg gccagcctgc tgttcgtggt catgtttatc ccagagacaa agggctgctc
     2341 cctggagcag atctctatgg agctggccaa ggtgaactac gtgaagaaca acatctgttt
     2401 catgagccac caccaggagg agctggtgcc caagcagcct cagaagcgga agccacagga
     2461 gcagctgctg gagtgcaaca agctgtgcgg aaggggacag tctaggcagc tgagccccga
     2521 gaccgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2581 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2641 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2701 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2761 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2821 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2881 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2941 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3001 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3061 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3121 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3181 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3241 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3301 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3361 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3421 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3481 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3541 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3601 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3661 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4081 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4141 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4201 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4261 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4321 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4381 ctggcctttt gctcacatgt t
//
